Comparison of the transcriptome, lipidome, and c-di-GMP production between BCG?BCG1419c and BCG, with Mincle- and Myd88-dependent induction of proinflammatory cytokines in murine macrophages.
Publication Title
Sci Rep
Document Type
Article
Publication Date
5-24-2024
Keywords
washington; isb
Abstract
We have previously reported the transcriptomic and lipidomic profile of the first-generation, hygromycin-resistant (HygR) version of the BCG?BCG1419c vaccine candidate, under biofilm conditions. We recently constructed and characterized the efficacy, safety, whole genome sequence, and proteomic profile of a second-generation version of BCG?BCG1419c, a strain lacking the BCG1419c gene and devoid of antibiotic markers. Here, we compared the antibiotic-less BCG?BCG1419c with BCG. We assessed their colonial and ultrastructural morphology, biofilm, c-di-GMP production in vitro, as well as their transcriptomic and lipidomic profiles, including their capacity to activate macrophages via Mincle and Myd88. Our results show that BCG?BCG1419c colonial and ultrastructural morphology, c-di-GMP, and biofilm production differed from parental BCG, whereas we found no significant changes in its lipidomic profile either in biofilm or planktonic growth conditions. Transcriptomic profiling suggests changes in BCG?BCG1419c cell wall and showed reduced transcription of some members of the DosR, MtrA, and ArgR regulons. Finally, induction of TNF-a, IL-6 or G-CSF by bone-marrow derived macrophages infected with either BCG?BCG1419c or BCG required Mincle and Myd88. Our results confirm that some differences already found to occur in HygR BCG?BCG1419c compared with BCG are maintained in the antibiotic-less version of this vaccine candidate except changes in production of PDIM. Comparison with previous characterizations conducted by OMICs show that some differences observed in BCG?BCG1419c compared with BCG are maintained whereas others are dependent on the growth condition employed to culture them.
Specialty/Research Institute
Institute for Systems Biology
DOI
10.1038/s41598-024-61815-8
